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PCR prevalence of rodent-‐borne Ljungan virus across Europe CrisPna Fevola1, 2,Chiara Rossi1, Annapaola Rizzoli1, Heikki HenTonen3, AnX Vaheri2, Anne Jääskeläinen2, 4, Edenext Collaborators*, Heidi C. Hauﬀe1 1: Department of Biodiversity and Molecular Ecology, Research and InnovaPon Centre, Fondazione Edmund Mach, San Michele All'Adige (TN) -‐ Italy 2: Department of Virology, University of Helsinki, Helsinki -‐ Finland 3: Finnish Forest Research InsPtute, Vantaa -‐ Finland 4: Helsinki University Central Hospital, Department of Virology and Immunology, Helsinki -‐ Finland *: D Arnoldi, M Collini, F Rosso, V Tagliapietra: FEM, IT; N Ferrari, C Romeo: Univ. Milan, IT; L Wauters: Univ. Insubria, IT; A Konecny: Masaryk Univ, CZ; J Piálek: ASCR, CZ; A Obiegala, C Silaghi: LMU, DE; S Fischer, R Ulrich: FLI, DE; N Charbonnel: INRA, FR;J Niemimaa, O VapalahP, L VouPlainen: FFRI, FI; T Sironen: UH, FI; C Reusken, JWB van der Giessen, M Maas: RIVM, NL; F Ecke, B Hörnfeldt, M Magnusson, Å Nordström, G Olsson, A Miller: SLU, SE; E Bužan: UP SRC, SL; M Kazimírová, M Stanko: SAS, SL. e-‐mail: cris3na.fevola@fmach.it The Ljungan (picorna) virus (LV) was idenPﬁed in 1998, in bank voles (Myodes glareolus, Figure 1) close to the Ljungan river, Sweden. Interest in LV has grown as a result of a suggested associaPon of the virus with some human pathologies. IntroducPon Figure 1: Myodes glareolus Since LV also causes disease in some wild rodents, it may have a role in small mammal cycles. This is the ﬁrst systemaPc screening of LV across the EU, especially in the bank vole, but also in other small mammals, including shrews and commensal species. Results Ø 1602 rodent liver samples screened by RT-‐PCR, including 24 species from 29 localiPes in 9 EU countries (14 sample sites did not have at least one LV-‐posiPve). Ø 167 samples were LV PCR-‐posiPve (Table 1). Table 1: List of the EU countries where animals were collected; number of samples screened in each country; number and prevalence of LV-‐posiPve samples. N. LV-‐posiPve Prevalence % N. screened (n. species (prevalence (n. species) posiPve) M. glareolus) Country Finland 278 (9) 20 (7) 7.19 (23.8) Sweden 441 (1)* 69 15.65 (15.65) Estonia 23 (1) 1 4.35 (n/a) Germany 103 (2)* 13 (1) 12.62 (13.0) Netherlands 30 (2) 1 (1) 3.33 (4.8) Slovakia 80 (1)* 3 3.75 (3.75) France 100 (1)* 11 11.0 (11.0) Italy 428 (14) 48 (7) 11.21 (26.2) CroaPa 119 (5) 1 (1) 0.84 (n/a) 1602 (24) 167 (13) 7.77 (10.91) The Figure 2: Map of the European countries where LV-‐posiPve animal samples were collected. = sampling sites where LV PCR-‐posiPve rodents where found. = sampling sites where LV has been found in previous studies. = sampling sites where all rodent samples were LV-‐negaPve. The land cover detecPon was not possible for Greece because it did not join the Corine land cover (CLC) project. Total Mean prevalence * indicates that the main or only species screened was the bank vole, M. glareolus. Conclusions LV-‐posiPve samples were found in all countries with signiﬁcant sample sizes, and in most species, including house mice, but not black rats. Overall PCR-‐prevalence in bank voles was about 11% (range 0-‐50% per populaPon). We added eight new species to the list of LV hosts (Fig. 3), including the red squirrel (Sciurus vulgaris), and a number of voles and shrews. Our study suggests that LV has a wide geographical and host distribuPon.
Materials and Methods As part of the EU FP7 project EDENext, rodent species from nine European countries were sampled. Using an LV-‐speciﬁc RT-‐PCR method (Donoso-‐Mantke et al., 2007), 1602 liver samples were screened for LV, including 831 bank voles. References red squirrel (Sciurus vulgaris) bicoloured shrew (Crocidura leucodon) valais shrew (Sorex an3norii) northern red-‐backed vole (Myodes ru3lus) grey red-‐backed vole (Myodes rufocanus) Acknowledgments The project was funded by EU grant FP7 261504 EDENext as well as Niklasson B et al. 1999. Virol 255:86-‐93; Jääskeläinen the FIRST PhD School and the Fondazione E. Mach. CF received a AJ et al. 2013. J Med Virol 85:2001-‐8; Hauﬀe HC et al. scholarship from CIMO (Finnish Ministry of EducaPon and Culture) 2010. J Wildl Dis 46:262-‐6; Donoso Mantke O et al. and a travel bursary from IRSAE (Research Council of Norway). 2007. J Virol Methods 141:171-‐7. Special thanks to MaTeo Marcantonio for creaPng the map. short-‐tailed vole lesser shrew (Crocidura suaveolens) (Microtus agres3s) tundra vole (Microtus oeconomus) Figure 3: new host species for LV.